Distinct CD16a features on human NK cells observed by flow cytometry correlate with increased ADCC.
- Resource Type
- Academic Journal
- Authors
- Benavente MCR; Department of Biochemistry and Molecular Biology, University of Georgia, 120 E. Green St., 30602, Athens, GA, Georgia.; Hakeem ZA; Department of Biochemistry and Molecular Biology, University of Georgia, 120 E. Green St., 30602, Athens, GA, Georgia.; Davis AR; Department of Biochemistry and Molecular Biology, University of Georgia, 120 E. Green St., 30602, Athens, GA, Georgia.; Murray NB; Complex Carbohydrate Research Center, University of Georgia, Athens, GA, Georgia.; Azadi P; Department of Biochemistry and Molecular Biology, University of Georgia, 120 E. Green St., 30602, Athens, GA, Georgia.; Complex Carbohydrate Research Center, University of Georgia, Athens, GA, Georgia.; Mace EM; Department of Pediatrics, Columbia University Irving Medical Center, New York, NY, USA.; Barb AW; Department of Biochemistry and Molecular Biology, University of Georgia, 120 E. Green St., 30602, Athens, GA, Georgia. abarb@uga.edu.; Complex Carbohydrate Research Center, University of Georgia, Athens, GA, Georgia. abarb@uga.edu.; Department of Chemistry, University of Georgia, Athens, GA, Georgia. abarb@uga.edu.
- Source
- Publisher: Nature Publishing Group Country of Publication: England NLM ID: 101563288 Publication Model: Electronic Cited Medium: Internet ISSN: 2045-2322 (Electronic) Linking ISSN: 20452322 NLM ISO Abbreviation: Sci Rep Subsets: MEDLINE
- Subject
- Language
- English
Natural killer (NK) cells destroy tissue that have been opsonized with antibodies. Strategies to generate or identify cells with increased potency are expected to enhance NK cell-based immunotherapies. We previously generated NK cells with increased antibody-dependent cell mediated cytotoxicity (ADCC) following treatment with kifunensine, an inhibitor targeting mannosidases early in the N-glycan processing pathway. Kifunensine treatment also increased the antibody-binding affinity of Fc γ receptor IIIa/CD16a. Here we demonstrate that inhibiting NK cell N-glycan processing increased ADCC. We reduced N-glycan processing with the CRIPSR-CAS9 knockdown of MGAT1, another early-stage N-glycan processing enzyme, and showed that these cells likewise increased antibody binding affinity and ADCC. These experiments led to the observation that NK cells with diminished N-glycan processing capability also revealed a clear phenotype in flow cytometry experiments using the B73.1 and 3G8 antibodies binding two distinct CD16a epitopes. We evaluated this "affinity profiling" approach using primary NK cells and identified a distinct shift and differentiated populations by flow cytometry that correlated with increased ADCC.
(© 2024. The Author(s).)